Combined therapy against tumors comprising substituted acryloyl distamycin derivatives, taxanes and/or antimetabolites

ABSTRACT

The present invention provides the combined use of acryloyl distamycin derivatives, in particular α-bromo- and -α-chloro-acryloyl distamycin derivatives of formula (I), as set forth in the specification, antimicrotubule agents and/or an antimetabolites. Also provided is the use of the said combination in the treatment or prevention of metastasis or in the treatment of tumors by inhibition of angiogenesis.

[0001] The present invention relates to the field of cancer treatmentand provides an antitumor composition comprising a substituted acryloyldistamycin derivative, more particularly an α-bromo- orα-chloro-acryloyl distamycin derivative, an antimicrotubule agent and/oran antimetabolite, having a synergistic antineoplastic effect.

[0002] Distamycin A and analogues thereof, hereinafter referred to asdistamycin and distamycin-like derivatives, are known in the art ascytotoxic agents useful in antitumor therapy.

[0003] Distamycin A is an antibiotic substance with antiviral andantiprotozoal activity, having a polypyrrole framework [Nature 203: 1064(1964); J. Med. Chem. 32: 774-778 (1989)]. The international patentapplications WO 90/11277, WO 98/04524, WO 98/21202, WO 99/50265, WO99/50266 and WO 01/40181 (claiming priority from British patentapplication No. 9928703.9), all in the name of the applicant itself andherewith incorporated by reference, disclose acryloyl distamycinderivatives wherein the amidino moiety of distamycin is optionallyreplaced by nitrogen-containing ending groups such as, for instance,cyanamidino, N-methylamidino, guanidino, carbamoyl, amidoxime, cyano andthe like, and/or wherein the polypyrrole framework of distamycin, orpart of it, is replaced by varying carbocyclic or heterocyclic moieties.

[0004] The present invention provides, in a first aspect, apharmaceutical composition for use in antineoplastic therapy in mammals,including humans, comprising a pharmaceutically acceptable carrier orexcipient;

[0005] an acryloyl distamycin derivative of formula (I):

[0006] wherein:

[0007] R₁ is a bromine or chlorine atom;

[0008] R₂ is a distamycin or distamycin-like framework; or apharmaceutically acceptable salt thereof; and

[0009] an antimicrotubule agent and/or an antimetabolite.

[0010] The present invention includes, within its scope, thepharmaceutical compositions comprising any of the possible isomerscovered by the compounds of formula (I), both considered separately orin admixture, as well as the metabolites and the pharmaceuticallyacceptable bio-precursors (otherwise known as pro-drugs) of thecompounds of formula (I).

[0011] In the present description, unless otherwise specified, with theterm distamycin or distamycin-like framework R₂ we intend any moietystructurally closely related to distamycin itself, for instance byoptionally replacing the ending amidino moiety of distamycin and/or itspolypyrrole framework, or part of it.

[0012] Antimicrotubule agents and antimetabolites are widely known inthe art as antitumor agents; see, for a general reference, Cancer,Principles and Practice of Oncology, Lippincott-Raven Ed. (1997),432-452 and 467-483

[0013] According to a preferred embodiment of the invention, herewithprovided are the above pharmaceutical compositions wherein theantimicrotubule agents are, for instance, taxanes, e.g. paclitaxel ordocetaxel; vinca alkaloids, e.g. vincristine, vinblastine, vindesine,vinorelbine; and estramustine, optionally encapsulated within liposomes.

[0014] Preferred antimetabolites are, for instance, antifolates, e.g.metotrexate, trimetrexate, tomudex; 5-fluoropyrimidines, e.g. 5-FU,floxuridine, ftorafur and capecitabine; cytidine analogs, e.g.cytarabine, azacitidine and gemcitabine.

[0015] Particularly preferred antimicrotubule agents are paclitaxel andestramustine whereas preferred antimetabolites are 5-fluorouracil orgemcitabine.

[0016] According to another preferred embodiment of the invention,herewith provided are the above pharmaceutical compositions wherein,within the acryloyl distamycin derivative of formula (I), R₁ has theabove reported meanings and R₂ is a group of formula (I) below:

[0017] wherein

[0018] m is an integer from 0 to 2;

[0019] n is an integer from 2 to 5;

[0020] r is 0 or 1;

[0021] X and Y are, the same or different and independently for eachheterocyclic ring, a nitrogen atom or a CH group;

[0022] G is phenylene, a 5 or 6 membered saturated or unsaturatedheterocyclic ring with from 1 to 3 heteroatoms selected among N, O or S,or it is a group of formula (III) below:

[0023] wherein Q is a nitrogen atom or a CH group and W is an oxygen orsulfur atom or it is a group NR₃ wherein R₃ is hydrogen or C₁-C₄ alkyl;

[0024] B is selected from the group consisting of

[0025] —CN; —NR₅R₆; —CONR₅R₆; —NHCONR₅R₆

[0026] wherein R₄ is cyano, amino, hydroxy or C₁-C₄ alkoxy; R₅, R₆ andR₇, the same or different, are hydrogen or C₁-C₄ alkyl.

[0027] In the present description, unless otherwise specified, with theterm C₁-C₄ alkyl or alkoxy group we intend a straight or branched groupselected from methyl ethyl, n-propyl isopropyl, n-butyl, isobutyl,sec-butyl, tert-butyl, methoxy, ethoxy, n-propoxy, isopropoxy, n-butoxy,isobutoxy, see-butoxy or tert-butoxy.

[0028] Even more preferred are the pharmaceutical compositions of theinvention comprising the above acryloyl distamycin derivative of formula(I) wherein R₁ is bromine or chlorine; R₂ is the above group of formula(II) wherein r is 0, m is 0 or 1, n is 4 and B has the above reportedmeanings.

[0029] Still more preferred, within this class, are the pharmaceuticalcompositions comprising the compounds of formula (I) wherein R₁ isbromine or chlorine; R₂ is the above group of formula (II) wherein r is0, m is or 1, n is 4, X and Y are both CH groups and B is selected from:

[0030] —CN; —CONR₅R₆; —NHCONR₅R₆

[0031] wherein R₄ is cyano or hydroxy and R₅, R₆ and R₇, the same ordifferent, are hydrogen or C₁-C₄ alkyl.

[0032] Pharmaceutically acceptable salts of the compounds of formula (I)are those with pharmaceutically acceptable inorganic or organic acidssuch as, for instance, hydrochloric, hydrobromic, sulfuric, nitric,acetic, propionic, succinic, malonic, citric, tararic, methanesulfonic,p-toluenesulfonic acid and the like.

[0033] Examples of preferred acryloyl distamycin derivatives of formula(I), within the compositions object of the invention, optionally in theform of pharmaceutically acceptable salts, preferably with hydrochloricacid, are:

[0034]1.N(5-{[(5-{[(5-{[(2-{[amino(imino)methyl]amino}ethyl)amino]carbonyl}-1-methyl-1H-pyrrol-3-yl)amino]carbonyl}-1-methyl-1H-pyrrol-3-yl)amino]carbonyl}-1-methyl-1H-pyrrol-3-yl)-4-[(2-bromoacryloyl)amino]-1methyl-1H-pyrrole-2-carboxamidehydrochloride;

[0035] 2.N(5-{[(5-{[(5-{[(2-{[amino(imino)methyl]amino}propyl)amino]carbonyl}-1-methyl-1H-pyrrol-3yl)amino]carbonyl}-1-methyl-1H-pyrrol-3-yl)amino]carbonyl}-1-methyl-1H-pyrrol-3-yl)-4-[(2-bromoacryloyl)amino]-1-methyl-1H-pyrrole-2-carboxamidehydrochloride;

[0036] 3.N-5-{[(5-{[(5-{[(3-amino-3-iminopropyl)amino]carbonyl}-1-methyl-1H-pyrrol-3-yl)amino]carbonyl}-1-methyl-1H-pyrrol-3yl)amino]carbonyl}-1-methyl-1H-pyrrol-3-yl)-4-[(2-bromoacryloyl)amino]-1-methyl-1H-pyrrole-2-carboxamidehydrochloride;

[0037] 4.N-(5-{[(5-{[(5-{[(3-amino-3-iminopropyl)amino]carbonyl}-1-methyl-1H-pyrrol-3-yl)amino]carbonyl}-1-methyl-1H-pyrrol-3-yl)amino]carbonyl}-1-methyl-1H-pyrrol-3-yl)-4-[(2-bromoacryloyl)amino]-1-methyl-1H-imidazole-2-carboxamidehydrochloride;

[0038]5.N-(5-{[(5-{[(5-{[(3-amino-3-iminopropyl)amino]carbonyl}-1-methyl-1H-pyrrol-3-yl)amino]carbonyl}-1-methyl-1H-pyrrol-3-yl)amino]carbonyl}-1-methyl-1H-pyrrol-3-yl)-3-[(2-bromoacryloyl)amino]-1-methyl-1H-pyrazole-5-carboxamidehydrochloride;

[0039] 6.N-(5-{[(5-{[(5-{[(3-amino-3-oxopropyl)amino]carbonyl}-1-methyl-1H-pyrrol-3-yl)amino]carbonyl}-1-methyl-1H-pyrrol-3-yl)amino]carbonyl}-1-methyl-1H-pyrrol-3-yl)-3-[(2-bromoacryloyl)amino]-1-methyl-1H-pyrazole-5-carboxamide;

[0040] 7.N-(5-{[(5-{[(5-{[(2-{[amino(imino)methyl]amino}ethyl)amino]carbonyl}-1-methyl-1H-pyrrol-3-yl)amino]carbonyl}-1methyl-1H-pyrrol-3-yl)amino]carbonyl}-1-methyl-1H-pyrrol-3-yl)-4-[(2-chloroacryloyl)amino]-1-methyl-1H-pyrrole-2-carboxamidehydrochloride;

[0041] 8.N-(5-{[(5-{[(3-{[amino(imino)methyl]amino}propyl)amino]carbonyl}-1-methyl-1H-pyrrol-3-yl)amino]carbonyl}-1-methyl-1H-pyrrol-3-yl)-4-[(2-bromoacryloyl)amino]-1-methyl-1H-pyrrole-2-carboxamidehydrochloride;

[0042] 9.N-5-{[(5-{[(3-amino-3-iminopropyl)amino]carbonyl}-1-methyl-1H-pyrrol-3-yl)amino]carbonyl}-1-methyl-1H-pyrrol-3-yl)-4-[(2-bromoacryloyl)amino]-1-methyl-1H-pyrrole-2-carboxamidehydrochloride; and

[0043] 10.N-{5-[({5-[({5-[({3-[(aminocarbonyl)amino]propyl}amino)carbonyl]-1-methyl-1H-pyrrol-3-yl}amino)carbonyl]-1-methyl-1H-pyrrol-3-yl}amino)carbonyl]-1-methyl-1H-pyrrol-3-yl}-4-[(2-bromoacryloyl)amino]-1-methyl-1H-pyyrole-2-carboxamide.

[0044] The above compounds of formula (I), either specificallyidentified as such or by means of the general formula, are known oreasily prepared according to known methods as reported, for instance, inthe aforementioned international patent applications WO 90/11277, WO98/04524, WO 98/21202, WO 99/50265 and WO 99/50266 and WO 01/40181.

[0045] The present invention further provides a product comprising anacryloyl distamycin derivative of formula (I), as defined above, anantimicrotubule agent and/or an antimetabolite, as a combinedpreparation for simultaneous, separate or sequential use in antitumortherapy.

[0046] Particularly preferred, in this respect, is a product comprisingN-(5-{[(5-{[(5-{[(2-[amino(imino)methyl]amino}ethyl)amino]carbonyl}-1-methyl-1H-pyrrol-3-yl)amino]carbonyl}-1-methyl-1H-pyrrol-3-yl)amino]carbonyl}-1-methyl-1H-pyrrol-3-yl)-4-[(2-bromoacryloyl)amino]-1-methyl-1H-pyrrole-2-carboxamidehydrochloride (internal code PNU 166196) and gemcitabine, as a combinedpreparation for simultaneous, separate or sequential use in antitumortherapy.

[0047] A further aspect of the present invention is to provide a methodof treating a mammal, including humans, suffering from a neoplasticdisease state, which method comprises administering to said mammal theabove acryloyl distamycin derivative of formula (I), an antimicrotubuleagent and/or an antimetabolite, in amounts effective to produce asynergistic antineoplastic effect.

[0048] The present invention also provides a method for lowering theside effects caused by antineoplastic therapy with an antineoplasticagent in a mammal in need thereof, including human, the methodcomprising administering to said mammal a combined preparationcomprising an acryloyl distamycin derivative of formula (I), anantimicrotubule agent and/or an antimetabolite, in amounts effective toproduce a synergistic antineoplastic effect.

[0049] By the term “synergistic antineoplastic effect”, as used herein,it is meant the inhibition of the growth tumor, preferably the completeregression of the tumor, by administering an effective amount of thecombination comprising an acryloyl distamycin derivative of formula (I),an antimicrotubule agent and/or an antimetabolite to mammals, includinghumans.

[0050] By the term “administered” or “administering”, as used herein, itis meant parenteral and/or oral administration; the term “parenteral”means intravenous, subcutaneous and intramuscular administration.

[0051] In the method of the present invention, the acryloyl distamycinderivative may be administered simultaneously with the antimicrotubuleagent or with the antimetabolite. Alternatively, the two drugs maybeadministered sequentially in either order.

[0052] When the acryloyl distamycin derivative is administered with boththe antimicrotubule agent and the antimetabolite, according to anembodiment of the invention, the drugs are preferably administeredsequentially, in any order.

[0053] In this respect, it will be appreciated that the actual preferredmethod and order of administration will vary according to, inter alias,the particular formulation of the acryloyl distamycin of formula (I)being used, the particular formulation of the antimicrotubule agentand/or the antimetabolite being used, the particular tumor model beingtreated as well as the particular host being treated.

[0054] To administer the acryloyl distamycin derivative of formula (I),according to the method of the invention, the course of therapygenerally employed comprises doses varying from about 0.05 to about 100mg/m² of body surface area and, more preferably, from about 0.1 to about50 mg/m² of body surface area.

[0055] For the administration of the taxanes, according to the method ofthe invention, the course of therapy generally employed comprises dosesvarying from about 1 to about 1000 mg/m² of body surface area and, morepreferably, from about 10 to about 500 mg/m² of body surface area.

[0056] For the administration of the vinca alkaloids, according to themethod of the invention, the course of therapy generally employedcomprises doses varying from about 0.1 to about 1000 mg/m² of bodysurface area and, more preferably, from about 0.5 to about 100 mg/m² ofbody surface area.

[0057] For the administration of the antimetabolite according to theinvention, the course of therapy generally employed comprises dosesvarying from about 0.1 to about 10 g/m² of body surface area and, morepreferably, from about 1 to about 5 g/m² of body surface area.

[0058] The antineoplastic therapy of the present invention isparticularly suitable for treating breast, ovary, lung, colon, kidney,stomach, pancreas, liver, melanoma, leukemia and brain tumors inmammals, including humans.

[0059] In a further aspect, the present invention is directed to acomposition comprising an effective amount of an acryloyl distamycinderivative of formula (I), as defined above, an antimicrotubule agentand/or an antimetabolite, in the preparation of a medicament for use inthe prevention or treatment of metastasis or in the treatment of tumorsby inhibition of angiogenesis.

[0060] As stated above, the effect of an acryloyl distamycin derivativeof formula (I) with an antimicrotubule agent and/or an antimetabolite,is significantly increased without a parallel increase of toxicity. Inother words, the combined therapy of the present invention enhances theantitumoral effects of the acryloyl distamycin derivative and of theother drug, being either an antimicrotubule, an antimetabolite or acombination thereof and, hence, provides the most effective and leasttoxic treatment for tumors.

[0061] The superadditive effects of the combined preparations of theinvention are shown, for instance, by the following in vivo antitumoractivity data which are intended to illustrate the present inventionwithout posing any limitation to it.

[0062] Table 1 shows the antileukemic activity on disseminatedL1210murine leukemia obtained by combiningN-5-{[(5-{[(5-{[(2-{[amino(imino)methyl]amino}ethyl)amino]carbonyl}-1-methyl-1H-pyrrol-3-yl)amino]carbonyl}-1-methyl-1H-pyrrol-3-yl)amino]carbonyl}-1-methyl-1H-pyrrol-3-yl)-4-[(2-bromoacryloyl)amino]-1-methyl-1H-pyrrole-2-carboxamidehydrochloride, as a representative compound of formula (I)—internal codePNU 166196, with gemcitabine.

[0063] At the dose of 15 mg/kg of gemcitabine alone (day +1 after tumorinjection, 2 h after PNU 166196 administration) and at the dose of 0.7mg/kg of PNU 166196 alone (days +1,6) were associated, without toxicity,ILS % values of 50 and 58, respectively. Combining gemcitabine and PNU166196 at the same doses with the same schedule, an increase of activitywith ILS % values of 127 were observed, thus indicating a synergisticeffect. TABLE 1 Antileukemic activity against disseminated L1210¹ murineleukemia of an acryloyl distamycin derivative (I) in combination withgemcitabine. Treatment Dose² Compound schedule (mg/kg/day) ILS %³ Tox⁴PNU 166196 iv +1.6 0.78 58 0/10 Gemcitabine iv +1 (*) 15 50 0/10 PNU166196 + iv +1.6 0.78+ 127 0/10 Gemcitabine iv +1 (*) 15

1. A pharmaceutical composition comprising a pharmaceutically acceptablecarrier or excipient and, as active ingredient, an acryloyl distamycinderivative of formula (I):

wherein: R₁ is a bromine or chlorine atom; R₂ is a distamycin ordistamycin-like framework; or a pharmaceutically acceptable saltthereof; and an antimicrotubule agent and/or an antimetabolite.
 2. Apharmaceutical composition according to claim 1 wherein theantimicrotubule agent is selected from taxanes, including paclitaxel anddocetaxel; vinca alkaloids, including vincristine, vinblastine,vindesine, vinorelbine; and estramustine, optionally encapsulated withinliposomes.
 3. A pharmaceutical composition according to claim 2 whereinthe antimicrotubule agent is paclitaxel or estramustine.
 4. Apharmaceutical composition according to claim 1 wherein theantimetabolite is selected from antifolates, including methotrexate,tomudex and trimetrexate; 5-fluoropyrimidine derivatives, including5-fluorouracil, floxuridine, ftorafur and capecitabine; and cytidineanalogs, including cytarabine, azacitidine and gemcitabine.
 5. Apharmaceutical composition according to claim 4 wherein theantimetabolite is selected from 5-fluorouracil or gemcitabine.
 6. Apharmaceutical composition according to claim 1 comprising an acryloyldistamycin derivative of formula (I)

wherein: R₁ is a bromine or chlorine atom; R₂ is a group of formula (II)

wherein m is an integer from 0 to 2; n is an integer from 2 to 5; r is 0or 1; X and Y are, the same or different and independently for eachheterocyclic ring, a nitrogen atom or a CH group; G is phenylene, a 5 or6 membered saturated or unsaturated heterocyclic ring with from 1 to 3heteroatoms selected among N, O or S, or it is a group of formula (III)below:

wherein Q is a nitrogen atom or a CH group and W is an oxygen or sulfuratom or it is a group NR₃ wherein R₃ is hydrogen or C₁-C₄ alkyl; B isselected from the group consisting of

—CN; —N₅R₆; —CONR₅R₆; —NHCONR₅R₆ wherein R₄ is cyano, amino, hydroxy orC₁-C₄ alkoxy; R₅, R₆ and R₇, the same or different, are hydrogen orC₁-C₄ alkyl.
 7. A pharmaceutical composition according to claim 6comprising an acryloyl distamycin derivative of formula (I) wherein R₁,R₂ and B are as defined in claim 6, r is 0, m is 0 or 1 and n is
 4. 8. Apharmaceutical composition according to claim 7 comprising an acryloyldistamycin derivative of formula (I) wherein R₁ and R₂ are as defined inclaim 6, r is 0, m is 0 or 1, n is 4, X and Y are both CH groups and Bis selected from:

—CN; —CONR₅R₆; —NHCONR₅R₆ wherein R₄ is cyano or hydroxy and R₅, R₆ andR₇, the same or different, are hydrogen or C₁-C₄ alkyl.
 9. Apharmaceutical composition according to claim 1 comprising an acryloyldistamycin derivative, optionally in the form of a pharmaceuticallyacceptable salt, selected from the group consisting of: 1.N(5-{[(5-{[(5-{[(2-{[amino(imino)methyl]amino}ethyl)amino]carbonyl}-1-methyl-1H-pyrrol-3-yl)amino]carbonyl}-1-methyl-1H-pyrrol-3-yl)amino]carbonyl}-1-methyl-1H-pyrrol-3-yl)-4-[(2-bromoacryloyl)amino]-1methyl-1H-pyrrole-2-carboxamidehydrochloride; 2.N(5-{[(5-{[(5-{[(2-{[amino(imino)methyl]amino}propyl)amino]carbonyl}-1-methyl-1H-pyrrol-3yl)amino]carbonyl}-1-methyl-1H-pyrrol-3-yl)amino]carbonyl}-1-methyl-1H-pyrrol-3-yl)-4-[(2-bromoacryloyl)amino]-1-methyl-1H-pyrrole-2-carboxamidehydrochloride; 3.N-5-{[(5-{[(5-{[(3-amino-3-iminopropyl)amino]carbonyl}-1-methyl-1H-pyrrol-3-yl)amino]carbonyl}-1-methyl-1H-pyrrol-3yl)amino]carbonyl}-1-methyl-1H-pyrrol-3-yl)-4-[(2-bromoacryloyl)amino]-1-methyl-1H-pyrrole-2-carboxamidehydrochloride; 4.N-(5-{[(5-{[(5-{[(3-amino-3-iminopropyl)amino]carbonyl}-1-methyl-1H-pyrrol-3-yl)amino]carbonyl}-1-methyl-1H-pyrrol-3-yl)amino]carbonyl}-1-methyl-1H-pyrrol-3-yl)-4-[(2-bromoacryloyl)amino]-1-methyl-1H-imidazole-2-carboxamidehydrochloride;5.N-(5-{[(5-{[(5-{[(3-amino-3-iminopropyl)amino]carbonyl}-1-methyl-1H-pyrrol-3-yl)amino]carbonyl}-1-methyl-1H-pyrrol-3-yl)amino]carbonyl}-1-methyl-1H-pyrrol-3-yl)-3-[(2-bromoacryloyl)amino]-1-methyl-1H-pyrazole-5-carboxamidehydrochloride; 6.N-(5-{[(5-{[(5-{[(3-amino-3-oxopropyl)amino]carbonyl}-1-methyl-1H-pyrrol-3-yl)amino]carbonyl}-1-methyl-1H-pyrrol-3-yl)amino]carbonyl}-1-methyl-1H-pyrrol-3-yl)-3-[(2-bromoacryloyl)amino]-1-methyl-1H-pyrazole-5-carboxamide;7.N-(5-{[(5-{[(5-{[(2-{[amino(imino)methyl]amino}ethyl)amino]carbonyl}-1-methyl-1H-pyrrol-3-yl)amino]carbonyl}-1methyl-1H-pyrrol-3-yl)amino]carbonyl}-1-methyl-1H-pyrrol-3-yl)-4-[(2-chloroacryloyl)amino]-1-methyl-1H-pyrrole-2-carboxamidehydrochloride; 8.N-(5-{[(5-{[(3-{[amino(imino)methyl]amino}propyl)amino]carbonyl}-1-methyl-1H-pyrrol-3-yl)amino]carbonyl}-1-methyl-1H-pyrrol-3-yl)-4-[(2-bromoacryloyl)amino]-1-methyl-1H-pyrrole-2-carboxamidehydrochloride; 9.N-5-{[(5-{[(3-amino-3-iminopropyl)amino]carbonyl}-1-methyl-1H-pyrrol-3-yl)amino]carbonyl}-1-methyl-1H-pyrrol-3-yl)-4-[(2-bromoacryloyl)amino]-1-methyl-1H-pyrrole-2-carboxamidehydrochloride; and 10.N-{5-[({5-[({5-[({3-[(aminocarbonyl)amino]propyl}amino)carbonyl]-1-methyl-1H-pyrrol-3-yl}amino)carbonyl]-1-methyl-1H-pyrrol-3-yl}amino)carbonyl]-1-methyl-1H-pyrrol-3-yl}-4-[(2-bromoacryloyl)amino]-1-methyl-1H-pyyrole-2-carboxamide.10. Products comprising an acryloyl distamycin derivative of formula(I):

wherein: R₁ is a bromine or chlorine atom; R₂ is a distamycin ordistamycin-like framework; or a pharmaceutically acceptable saltthereof, an antimicrotubule agent and/or an antimetabolite, as acombined preparation for simultaneous, separate or sequential use in thetreatment of tumors.
 11. Products according to claim 10 wherein theantimicrotubule agent is selected from taxanes, including paclitaxel anddocetaxel; vinca alkaloids, including vincristine, vinblastine,vindesine, vinorelbine; and estramustine, optionally encapsulated withinliposomes.
 12. Products according to claim 11 wherein theantimicrotubule agent is paclitaxel or estramustine.
 13. Productsaccording to claim 10 wherein the antimetabolite is selected fromantifolates, including methotrexate, tomudex and trimetrexate;5-fluoropyrimidine derivatives, including 5-fluorouracil, floxuridine,ftorafur and capecitabine; and cytidine logs, including cytarabine,azacitidine and gemcitabine.
 14. Products according to claim 13 whereinthe antimetabolite is selected from 5-fluorouracil or gemcitabine. 15.Products according to claim 10 comprising an acryloyl distamycinderivative of formula (I)

wherein: R₁ is a bromine or chlorine atom; R₂ is a group of formula (II)

wherein m is an integer from 0 to 2; n is an integer from 2 to 5; r is 0or 1; X and Y are, the same or different and independently for eachheterocyclic ring, a nitrogen atom or a CH group; G is phenylene, a 5 or6 membered saturated or unsaturated heterocyclic ring with from 1 to 3heteroatoms selected among N, O or S, or it is a group of formula (III)below:

wherein Q is a nitrogen atom or a CH group and W is an oxygen or sulfuratom or it is a group NR₃ wherein R₃ is hydrogen or C₁-C₄ alkyl; B isselected from the group consisting of

—CN; —NR₅R₆; CONR₅R₆; —NHCONR₅R₆ wherein R₄ is cyano, amino, hydroxy orC₁-C₄ alkoxy R₅, R₆ and R₇, the same or different, are hydrogen or C₁-C₄alkyl.
 16. Products according to claim 10 wherein the acryloyldistamycin derivative is selected from the group as defined in claim 9.17. Use of an acryloyl distamycin derivative of formula (I), as definedin claim 1 or in any one of claims from 6 to 9 in the preparation of amedicament for use in combination therapy with an antimicrotubule agentand/or an antimetabolite in the treatment of tumors.
 18. Use accordingto claim 17 wherein the medicament further comprises the saidantimicrotubule agent and/or antimetabolite.
 19. Use according to claim17 or 18 wherein the acryloyl distamycin derivative is selected from thegroup as defined in claim
 9. 20. Use according to any one of claims 17to 19 wherein the tumor is selected from breast, ovary, lung, colon,kidney, stomach, pancreas, liver, melanoma, leukemia and brain tumors.21. Use of an acryloyl distamycin derivative of formula (I), as definedin claim 1 or in any one of claims form 6 to 9 in the preparation of amedicament for use in combination therapy with an antimicrotubule agentand/or an antimetabolite in the prevention or treatment of metastasis orin the treatment of tumors by inhibition of angiogenesis.
 22. Useaccording to claim 21 wherein the medicament further comprises the saidantimicrotubule agent and/or antimetabolite.
 23. A method of treating amammal including humans, suffering from a neoplastic disease state,which method comprises administering to said mammal the acryloyldistamycin derivative of formula (I), as defined in claim 1 or any oneof claims 6 to 9, an antimicrotubule agent and/or an, antimetabolite, inamounts effective to produce a synergistic antineoplastic effect.
 24. Amethod for lowering the side effects caused by antineoplastic therapywith an antineoplastic agent, in a mammal in need thereof includinghumans, the method comprising administering to said mammal a combinedpreparation comprising an antimicrotubule agent and/or an antimetabolitewith an acryloyl distamycin derivative of formula (I), as defined inclaim 1 or any one of claims from 6 to 7, in amounts effective toproduce a synergistic antineoplastic effect.
 25. A pharmaceuticalcomposition according to claim 1 wherein the acryloyl distamycinderivative, optionally in the form of a pharmaceutically acceptablesalt, isN-(5-{[(5-{[(5-{[(2-{[amino(imino)methyl]amino}ethyl)amino]carbonyl}-1-methyl-1H-pyrrol-3-yl)amino]carbonyl}-1-methyl-1H-pyrrol-3-yl)amino]carbonyl}-1-methyl-1H-pyrrol-3-yl[(2-bromoacyloyl)amino]-1-methyl-1H-pyrrole-2-carboxamide,and the antimetabolite is gemcitabine.